Novel processes have been discovered for the preparation of novel lecithin derived glycolipids and phospholipids. Those lecithin derived glycolipids and phospholipids and dilutions thereof, as well as lecithin dilutions, and the dosage unit forms which have been discovered are useful in the treatment of a wide range of diseases. These lecithin fractions and dilutions have superior purity and pharmacological capabilities. Methods of treatment of warm-blooded animals using either or both the products of these methods and/or specific lecithin dilutions have also been discovered.
Shinitzky et al., U.S. Pat. No. 4,474,773, disclose lipid extracts obtained from egg yolk or soybean which are said to be useful for treating various diseases and physiological conditions as well as a process for the fractionation of lipids. The fractionation is effected by dissolving a lipid extract in a suitable solvent, evaporating the solvent to almost complete dryness, precipitating a fraction of the dissolved lipids by the addition of an organic solvent, and recovering the desired fraction from the supernatant. Three preferred methods of fractionation are exemplified. The first method involves dissolving the lipid extract in chloroform, evaporating to near dryness, and precipitating by the addition of acetone. The supernatant is removed and is evaporated to near dryness, leaving a fraction of about 5 weight percent of the untreated extract as the active lipid. The second method mixes a natural lipid source with acetone to remove excess undesired lipids. The precipitate is treated again with acetone, and the supernatant is collected and is evaporated to complete dryness, leaving the desired fraction which is about 10 to 15 weight percent of the initial quantity of the lipid source. In the third method, a natural lipid source is mixed with acetone to remove excess undesired lipid. The precipitate is treated again with acetone, and the supernatant is collected and is cooled below 0.degree. C. at which time the active lipid (about 10 to 15 weight percent of the initial quantity of the lipid source) is precipitated and is collected.
Chang et al., U.S. Pat. No. 4,563,354, disclose alcohol-soluble fractions of vegetable lecithin which are suitable for use as emulsifiers for oil-in-water emulsions. These fractions are well-tolerated when used for parenteral nutrition. The Chang et al. extraction process includes a preliminary purification of the lecithin which involves dissolving the lecithin in a solvent such as hexane, and precipitating the lecithin through the use of acetone. The precipitated lecithin is then extracted with an alcohol at a temperature ranging from -20.degree. C. to slightly elevated temperatures in order to extract the alcohol-soluble portion of the lecithin. The extraction may be repeated multiple times. The filtrate(s) from the alcohol extraction step(s) are combined and are chilled to about -20.degree. C. for about 24 hours to produce a precipitate. Any solids which separate are removed by filtration and are discarded. The resulting filtrate is freed of solvent under vacuum and is again dissolved in alcohol. The resulting solution is chilled, again utilizing a temperature of about -20.degree. C. for about 24 hours. The precipitate is then removed by filtration and is discarded. The resulting filtrate again is treated under vacuum to remove solvent, and the residue is precipitated from acetone to yield the alcohol-soluble soybean phospholipid fraction.
U.K. Patent Publication No. 2,004,741 illustrates a composition which is said to be useful in treating baldness, burns and various viral complaints and which contains cholesterol, an unsaturated fatty acid, and a phosphoamino-lipid. The phosphoamino-lipid utilized in the composition may be egg lecithin or vegetable lecithin.
Japanese Patent Publication No. 55/50859 discloses a method for refining lecithin from, for example, soybean oil, by adding propylene glycol to extract the oil and using an aqueous organic acid to coagulate the lecithin. The extraction is practiced at below 80.degree.C.
Japanese Patent Publication No. 55/054861 also deals with a method for treating crude lecithin-containing oil by adding propylene glycol, mixing to extract the lecithin, adding a group II metal chloride, mixing, and separating the coagulated lecithin.
Japanese Patent Publication No. 62/246525 relates to a pharmacological lecithin mixture which is prepared by suspending phosphatidyl choline, phospholipid mixture and neutral lipid in an alcohol solvent, filtering, and removing the solvent. This lecithin mixture contains at least 75 weight percent phosphatidyl choline, 5 to 13 weight percent of an ethanol amine and phosphatidic acid-containing phospholipid mixture, and 0 to 20 weight percent of neutral lipid. The lecithin mixture is derived from animal or vegetable sources such as seeds, fruit, egg yolk, soybean, and the like and is useful as a medicine for arteriosclerosis, hyperlipemia, liver diseases, heart attack, and the like.
European Patent Application No. 0,213,724 relates to a lipid mixture for membrane fluidization consisting of about 7 parts by weight of neutral lipids and about 3 parts by weight of phospholipids. The phospholipids consist of phosphatidyl choline, phosphatidyl ethanol amine, and neutral lipids consisting essentially of glycerides.
European Patent Application No. 0,214,055 discloses an artificial surfactant which is a mixture of dipalmitoyl phosphatidyl choline, distearoyl phosphatidyl choline and soybean lecithin. Soybean lecithin is a well-known adjuvant for intravenous preparations, and the soybean lecithin utilized in EPO '055 has a molecular weight of about 780.
European Patent Application No. 0,239,729 relates to polyprenyl alcohol-containing injections comprising certain polyprenyl alcohols and lecithin as an essential ingredient. The addition of a lecithin increases activity of the polyprenyl alcohol in in vivo applications.
Tayot, U.S. Pat. No. 4,415,733, discloses ganglioside derivatives which are products of partial deacylation of gangliosides. Free amino groups are demonstrated by a positive reaction in a ninhydrin test, are mobile in chromatograph on a thin silica gel layer in the chloroform-butanol-water system (60:32:7), and exhibit specific affinity properties of the gangliosides from which they are derived. The derivatives are able to couple to solid supports by amino groups that have appeared during partial deacylation without loss of specific affinity properties. Tayot also relates to a process of preparing these activated ganglioside derivatives while retaining their biological properties. The activated gangliosides are treated with an aqueous base solution at a temperature between 0.degree. and 120.degree. C. The lower temperatures require more basic medium. The reaction time for partial deacylation varies from 30 minutes to 24 hours. The transformation is reflected by the appearance of at least an -NH.sub.2 function resulting from partial hydrolysis of the N-acetyl or N-acyl function in sufficient number to allow fixing by the known methods, but limited enough to maintain the biological properties of the gangliosides.
Alving et al., U.S. Pat. No. 4,416,872, disclose a method for the prophylaxis and the chemotherapeutic treatment of malaria with antimalarial compositions comprising either a glycolipid alone, a glycoprotein encapsulated within a liposome, or an 8-aminquinoline drug and a glycoconjugate contained within a liposome. The liposome (exclusive of the glycoconjugate constituent) may contain a combination of (1) a phospholipid and cholesterol or (2) a phospholipid, cholesterol and a negatively or positively charged (lipophilic) amphipathic compound. Glycolipids having a terminal glucose or galactose moiety are said to be effective when incorporated within the liposomes. The amphipathic component, by way of example, can be diacetyl phosphate or stearylamine to impart charge to the liposomes. Glycolipids having a terminal glucose or galactose moiety are said to be effective when incorporated within the liposomes.
Dingerdissen et al., U.S. Patent No. 4,694,069, disclose glycopeptide AAD-609 antibiotics which are structurally and biologically related to the AAD-216 antibiotics. The AAD-609 antibiotic varies from AAD-216 antibiotic in that the sugar moiety within the glycolipid radical is glucosamine rather than aminoglucuronic acid. Pharmaceutical compositions containing at least one of the AAD-609 factors and a pharmaceutically acceptable carrier are discussed as well. The compositions may additionally contain other active antibacterial agents.
Graves, U.S. Pat. No. 4,829,009, discloses matrix layers suitable for solid phase immunoassays useful in eliminating false positives. Graves provides a means for verification of the effectiveness of such matrix layers and for maximizing their effectiveness, as well as for quantitation of background noise. Generally, the noise reduction component will provide an anionic barrier and therefore will be a negatively charged material, preferably a macromolecule. Any negatively charged (at neutral pH) protein, glycoprotein, polyamino acid or other natural or synthetic macromolecule could be used as a noise reduction component. Graves also includes a method of designing a protocol to determine simultaneously the values of three parameters--sensitivity ratio, signal to noise ratio, and noise balance ratio--which collectively influence the matrix index.
Kojima et al., U.S. Pat. No. 4,871,540, disclose a process for producing a biologically active substance of plant origin, having interferon-inducing activity. The substance is said to be useful for preventing and treating, for example, aesthema, rhintitis, B-type hepatitis, AIDS and other allergic diseases and immunological insufficiencies. The active substance is said to be capable of inhibiting the formation of IgE antibodies, promoting the formation of IgG and IgM antibodies and having interferon inducing activity, polyclonal B lymphocyte activating activity, mitogenic activity, adjuvant activity and anti-tumor activity.
Hakomori, U.S. Pat. No. 4,876,199, relates to hybridoma cell lines that produce monoclonal antibodies which differentially recognize glycolipids with mono-, di- and trifucosylated type 2 chain structures. These monoclonal antibodies are said to be useful for detecting specific types of tumor cells in the diagnosis and the treatment of human cancer.
Stolze and Khaden, Institute of Virology, "Apparent Lack of Neutralizing Antibodies in Aleutran Diseases Is Due to Masking of Antigenic Sites by Phospholipids", Academic Press, Inc., 1987, discuss neutralization of Aleutian disease virus by phospholipids.
Nakashima et al. "Inhibitory Effect of Glycosides Like Saponin From Soybean on the Infectivity of HIV In Vitro, AIDS Vol. 3, p. 655-658 (1989), describe soybean saponins isolated from the soybean seeds which were investigated for their antiviral activity on HIV in vitro, using an HTLV-1-carring cell line, NT-4. One of the saponins (identified as saponin B1) completely inhibited HIV-induced cytopathic effects and virus-specific antigen expression six (6) days after infection at concentrations greater than 0.5 milligrams per milliliter. Another saponin (identified as saponin B2) also inhibited HIV infection, although less potently. Both saponins are reported to have had no directed effect on the reverse transcriptase activity of HIV. Saponin B1 was reported to have also inhibited HIV-induced cell fusion in the MOLT-4 cell system.
The present applicants believe that the activity described by Nakashima et al. is limited to in vitro activity. The present applicants have found that the in vivo activity of the presently described lecithin fractions and dilutions is dependent upon at least some phospholipid content residing in the lecithin fractions and/or dilutions.
The present applicants have found that as the content of phospholipid with its complexing agents is reduced, the in vivo activity is also reduced.